E. Calderon et al., MODULATION OF ENDOTHELIN-1 PRODUCTION BY A PULMONARY EPITHELIAL-CELL LINE .1. REGULATION BY GLUCOCORTICOIDS, Biochemical pharmacology, 48(11), 1994, pp. 2065-2071
Endothelin-1 (ET-1) is one of the most potent bronchoconstrictor agent
s yet described. Bronchial epithelial cells of asthmatic patients in v
ivo express preproET-1 and in vitro release high amounts of ET-1. Heal
thy and chronic bronchitic controls do not express preproET-1 or relea
se ET-1. Interleukin-2 (IL-2) and other cytokines up-regulate the in v
itro ET-1 release in guinea pig airway epithelial cells. We explored w
hether two glucocorticoids, dexamethasone (Dex) and triamcinolone acet
onide (TA), inhibit the synthesis and release of ET-1 by A549 cells, a
transformed human pulmonary epithelial cell line, since ET-1 may have
a basic role in the pathogenesis of asthma. Cells were grown to confl
uence in RPMI 1640 plus 10% fetal bovine serum (FBS). Cells were then
cultured for 3 days without serum to obtain ET-I basal levels. The eff
ects of 10% FBS, IL-2 (10 U/mL), Dex, TA or mifepristone, a steroid an
tagonist (1, 10 or 100 nM), were evaluated on ET-1 as measured by radi
oimmunoassay (RIA). ET-1 production increased from 57.6 +/- 5 pg/mg ce
ll protein at 6 hr to 170 +/- 9 pg/mg cell protein at 72 hr in control
cultures. Ten percent FBS increased ET-1 production from 58.7 +/- 9.6
to 399 +/- 14.5 pg/mg cell protein. IL-2 significantly increased ET-1
from 100.7 +/- 6.1 to 144 +/- 6.7 at 24 hr and from 170 +/- 9 to 207.
7 +/- 24 at 72 hr. Dex and TA (10 and 100 nM) at 24-72 hr decreased ET
-1 under basal conditions. Both drugs (only at 100 nM) decreased ET-1
production in 10% FBS- and IL-2-stimulated cells. Mifepristone (10 and
100 nM) reversed the decreased production of ET-1 induced by Dex (100
nM) at 24-72 hr. Northern blot analysis showed that Dex (100 nM) decr
eased the expression of ET-1 mRNA at 6 and 24 hr, but that mifepriston
e (100 nM) reversed this effect in cells cultured with Dex. In conclus
ion, Dex and TA down-regulate the synthesis and production of ET-1 by
this human pulmonary epithelial cell line under basal or stimulated co
nditions, and these effects are reversed by mifepristone. These findin
gs suggest a novel mechanism of glucocorticoid effect during the treat
ment of asthma.